The reagents were of analytical
grade unless stated otherwise. All analyses were performed on samples
taken from the aerated bioreactor unless stated otherwise. The liquid
was circulated through all sections of the system (i.e., H
2S absorber, anaerobic bioreactor and aerated bioreactor) with a flow
of 8 kg/h (streams 1, 2, and 3 in
Figure 1). Hence, the HRT’s in the different
sections of the process were: 4 min in the absorber bottom, 44 min
in the anaerobic bioreactor and 94 min in the aerated bioreactor.
Therefore, the alkalinity and concentrations of SO
42–, S
2O
32–,
S
8 and bacteria were equal throughout the complete system
(which was verified by measurements) and the concentrations measured
in samples of the aerated bioreactor were also representative for
the liquid in the H
2S absorber and anaerobic bioreactor.
The composition of the treated gas of the H
2S absorber
(sampled every 3 min) was analyzed for H
2S, CO
2 and N
2, using a gas chromatograph (GC) (EnCal 3000, Honeywell,
Morristown, NJ). The GC was equipped with a molsieve and a ppu column
(both 10m), using helium as carrier gas. The molsieve column was operated
at a pressure of 200 kpa and 100 °C. The GC was calibrated weekly.
To determine the HS
– removal in the anaerobic
bioreactor, the total sulfide concentration (
Stot2–), which is the sum of S
2–, HS
– and polysulfide-sulfane (S
x2–), was measured in a sample of the anaerobic
reactor by titration with a solution of 0.1 M AgNO
3, using
a Titrino Plus Titrator (Metrohm, Herisau, Switzerland). Before titration,
the tested sample was filtered over a 0.45 μm cellulose acetate
membrane filter to remove S
8 and bacteria. Two mL of filtered
sample was added to 80 mL 4% (w/v) NaOH, with 1 mL of 30% (w/v) NH
4OH to stabilize
Stot2–. A comparison between unfiltered and filtered samples did not show
significant differences.
The specific HS
– removal
efficiency in the anaerobic
bioreactor (mg S/mg N) was calculated based on the H
2S
load, the liquid flows, the measured HS
– concentration
and the biomass concentration, according to
eq 1. Here,
H2S load
is the mass loading in the H
2S absorber
(mg S h
–1), and the solution flow is the liquid
flow to the anaerobic reactor (L h
–1). [
Stot2–]
meas is the total measured sulfide concentration (mg
S L
–1) and Xb is the biomass concentration (mg N
L
–1).
The biomass concentration was measured
as the amount of total organic
N using the Dr. Lange cuvette test LCK138 (Hach Lange, Germany). The
difference between the supernatant (i.e., a sample centrifuged for
10 min at 14 000
g) and a noncentrifuged sample
indicated the total amount of N present in the biomass. It was confirmed
that the presence of biologically produced S
8 did not affect
the N analyses, provided that the samples were at least 5 times diluted.
Considering the generic stoichiometric chemical equation for HA-SOB,
that is, CH
1.8O
0.5N
0.2,
20 (link) the total N amount accounts for 10 mol % of
the total dry weight biomass.
The specific conductivity of the
samples was monitored using an
offline conductivity sensor (LF 340, WTW, Weilheim, Germany). The
alkalinity was measured with titration with 0.1 M HCL to pH 4.3, using
a titrator (Titralab AT1000, Hach Lange, Germany). The method was
verified with an analysis in accordance with WAC/III/A/006.
After removal of cells and S
8 by centrifugation for
10 min at 14 000
g, the supernatant samples
were analyzed for COD—in duplicate, (Lange cuvette test LCK514,
Hach Lange Germany) at 605 nm, and using a spectrophotometer (Hach
Lange, Germany)—to determine total dissolved S
2O
32–. In addition, the sample was analyzed
for SO
42–, by using a Hach Lange cuvette
test LCK353 and spectrophotometer at 800 nm. These methods were verified
with an ion chromatography method, in accordance with ISO 10304-1.
The concentration of total suspended solids (TSS), consisting of
mainly S
8, was analyzed in triplicate. A 5–15 mL
sample (depending on the TSS concentration) was filtered over a predried
(24h, 60 °C) and preweighed GF/C Glass microfiber filter (Whatman).
After drying (60 °C for at least 24h), the filters were weighed
again. The TSS was determined as the difference between the final
weight and initial weight, divided by the sample weight.
As
the formed S
8 particles have a tendency to attach
to the reactor wall, it was not possible to calculate the S
8 production rate from the analyses. As no products other than S
8, SO
42– and S
2O
32– were measured in the reactor,
7 (link) the production rate of S
8 could be
calculated from the following mass balance: Here,
PS8,
PSO42– and
PS2O32– are the production
rates of S
8,
SO
42–, and S
2O
32–, respectively, in mol S-product day
–1 and I
H2S is the volumetric H
2S influent in
mol S day
–1. The production rates of both SO
42– and S
2O
32– (not shown) are calculated as follows: The selectivities were calculated
according to
eq 4: Here, effluent is
the total
effluent of the system (L) in time interval Δ
t (days) (i.e., sample volumes, S
8 slurry, and bleed),
[ ] the average concentration (mol
S L
–1) over time interval Δ
t,
V the total liquid volume of the system (18.7
L) and Δ[ ] the concentration
changes (mol S L
–1) over time interval Δ
t. The
mathematical equation to determine the caustic use and bleed flow
of a full scale system based on the product selectivities can be found
in the
Supporting Information (SI 4).
The samples for microbial community analysis were conserved immediately
after sampling by addition of ethanol up to 50% (v/v). DNA was extracted
with the MPbio FastDNA SPIN Kit for Soil. Subsequently, PCR was used
to amplify the V3 and V4 region of the 16S rRNA gene of bacteria giving
a 400bp product. The library prep, sequencing and data analysis was
performed via the 16S BioProphyler method,
21 using the Illumina PE300 platform and MiSeq sequencer. Within the
BioProphyler method all primer sequences and low quality reads were
deleted from the raw data sets. The obtained sequences were compared
with the online nt database with the aid of the BLAST algorithm. Low
abundance reads were not removed from the data set and no correction
on differences in library size was applied. The reported species name
is the species which is most related to the detected sequence. The
Supporting Information contains the accession
numbers of the sequences with the respective number of reads. The
EMBL-EBI accession number for presented 16S rRNA sequencing set is
PRJEB31478.
de Rink R., Klok J.B., van Heeringen G.J., Sorokin D.Y., ter Heijne A., Zeijlmaker R., Mos Y.M., de Wilde V., Keesman K.J, & Buisman C.J. (2019). Increasing the Selectivity for Sulfur Formation in Biological Gas Desulfurization. Environmental Science & Technology, 53(8), 4519-4527.